Engineering a Photoautotrophic Microbial Coculture toward Enhanced Biohydrogen Production

The application of synthetic phototrophic microbial consortia holds promise for sustainable bioenergy production. Nevertheless, strategies for the efficient construction and regulation of such consortia remain challenging. Applying tools of genetic engineering, this study successfully constructed a synthetic community of phototrophs using <i>Rhodopseudomonas palustris</i> (<i>R. palustris</i>) and an engineered strain of <i>Synechocystis sp</i> PCC6803 for acetate production (<i>Synechocystis_acs</i>), enabling the production of biohydrogen and fatty acids during nitrogen and carbon dioxide fixation. Elemental balance confirmed carbon capture and nitrogen fixation into the consortium. The strategy of circadian illumination effectively limited oxygen levels in the system, ensuring the activity of the nitrogenase in <i>R. palustris</i>, despite oxygenic photosynthesis happening in <i>Synechocystis</i>. When infrared light was introduced into the circadian illumination, the production of H₂ (9.70 μmol mg^-1) and fatty acids (especially C16 and C18) was significantly enhanced. Proteomic analysis indicated acetate exchange and light-dependent regulation of metabolic activities. Infrared illumination significantly stimulated the expression of proteins coding for nitrogen fixation, carbohydrate metabolism, and transporters in <i>R. palustris</i>, while constant white light led to the most upregulation of photosynthesis-related proteins in <i>Synechocystis_acs</i>. This study demonstrated the successful construction and light regulation of a phototrophic community, enabling H₂ and fatty acid production through carbon and nitrogen fixation.